Electrical field stimulation induces cardiac fibroblast proliferation through the calcineurin-NFAT pathway
Electrical field stimulation induces cardiac fibroblast proliferation through the calcineurin-NFAT pathway
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DOI：
10.1139/y2012-133
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发表时间：
2012-12-01
影响因子：
2.1
通讯作者：
Zhu, Wei-Zhong
中科院分区：
文献类型：
--
作者：
Chen, Qing-Qing;Zhang, Wei;Zhu, Wei-Zhong
大多数心脏疾病都与纤维化有关。钙调磷酸酶（Calcineurin, CaN）受Ca2+/calmodulin （CaM）调控。CaN-NFAT（活化T细胞核因子）通路参与心脏肥大等心脏疾病的发生过程，但其在心肌纤维化中的作用尚不清楚。本研究探讨了CaN-NFAT通路是否参与了电场刺激（EFS）诱导的心脏成纤维细胞（CF）增殖，这是近年来治疗心力衰竭和心脏组织工程的一种流行方法。通过细胞存活试验（MTT）和细胞计数评估CF增殖。通过ⅰ型胶原蛋白和ⅲ型胶原蛋白的表达评估心肌纤维化程度。采用绿色荧光蛋白（GFP）标记的NFAT检测NFAT核易位。EFS可增强CF增殖、心肌纤维化、CaN活性和NFAT核易位。更重要的是，这些作用被CaN抑制剂、显性阴性CaN （DN-CaN）和用siRNA沉默的CaN基因所消除。此外，用BAPTA-AM缓冲细胞内Ca2+和用硝苯地平阻断Ca2+内流抑制efs诱导的细胞内Ca2+和CF增殖的增加。这些结果提示CaN-NFAT通路介导CF增殖，CaN-NFAT通路可能是efs诱导心肌纤维化和心脏组织工程的潜在治疗靶点。
Most cardiac diseases are associated with fibrosis. Calcineurin (CaN) is regulated by Ca2+/calmodulin (CaM). The CaN-NFAT (nuclear factor of activated T cell) pathway is involved in the process of cardiac diseases, such as cardiac hypertrophy, but its effect on myocardial fibrosis remains unclear. The present study investigates whether the CaN-NFAT pathway is involved in cardiac fibroblast (CF) proliferation induced by electrical field stimulation (EFS), which recently became a popular treatment for heart failure and cardiac tissue engineering. CF proliferation was evaluated by a cell survival assay (MTT) and cell counts. Myocardial fibrosis was assessed by collagen I and collagen III protein expression. Green fluorescent protein (GFP)-tagged NFAT was used to detect NFAT nuclear translocation. CF proliferation, myocardial fibrosis, CaN activity, and NFAT nuclear translocation were enhanced by EFS. More importantly, these effects were abolished by CaN inhibitors, dominant negative CaN (DN-CaN), and CaN gene silenced with siRNA. Furthermore, buffering intracellular Ca2+ with BAPTA-AM and blocking Ca2+ influx with nifedipine suppressed EFS-induced increase in intracellular Ca2+ and CF proliferation. These results suggested that the CaN-NFAT pathway mediates CF proliferation, and that the CaN-NFAT pathway might be a possible therapeutic target for EFS-induced myocardial fibrosis and cardiac tissue engineering.